Surfactant Protein–C Chromatin-Bound Green Fluorescence Protein Reporter Mice Reveal Heterogeneity of Surfactant Protein C–Expressing Lung Cells

Kim, Jonghwan; Gludish, David W.; Roach, Rebecca R.; Saunders, Arven; Barrios, Juliana; Woo, Andrew; Chen, Huaiyong; Conner, David A.; Fujiwara, Yuko; Stripp, Barry R.; Kim, Carla F.

doi:10.1165/rcmb.2011-0403oc

Cited by 57 publications

(61 citation statements)

References 45 publications

(60 reference statements)

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“…This homogeneity demonstrates an important characteristic of our culture system: that we are starting with a well-characterized, specific population of cells known to initially reside in the alveoli that gives rise to a homogenous population of colonies containing only alveolar cells. This is in contrast to the populations of distal progenitor cells studied by other investigators that ultimately yield mixed colony types containing both alveolar and bronchiolar cells (25,37,38).…”

Section: Discussioncontrasting

confidence: 71%

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Type 2 alveolar cells are stem cells in adult lung

et al. 2013

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Section: Discussioncontrasting

confidence: 71%

“…This uniformity is in marked contrast to the multiple colony types described by other investigators in which distal lung cells, sorted by surface marker expression, are plated in 3D Matrigel assays (25,37,38).…”

Section: Single Lineage-labeled Aec2s Form 3d Alveolar-like Structurementioning

confidence: 65%

Type 2 alveolar cells are stem cells in adult lung

et al. 2013

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“…Because the existing methods for isolation of murine ATII cells yield preparations of approximately 85-90% pure cells, this method is a significant improvement (12). Alternative methods for isolation of pure murine ATII cells require use of transgenic green fluorescent protein (GFP) reporter mice driven by the ATIIspecific SP-C promoter, and even then tend to yield heterogeneous cell populations from adult animals (45). Our method is also the first described method to obtain high-purity mTEC preparations fresh from the mouse trachea.…”

Section: Discussionmentioning

confidence: 99%

Immune Defense Protein Expression in Highly Purified Mouse Lung Epithelial Cells

Sinha

Lowell

2016

Am J Respir Cell Mol Biol

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Lung epithelial cells play critical roles in initiating and modulating immune responses during pulmonary infection or injury. To better understand the spectrum of immune response-related proteins present in lung epithelial cells, we developed an improved method of isolating highly pure primary murine alveolar type (AT) II cells and murine tracheal epithelial cells (mTECs) using negative selection for a variety of lineage markers and positive selection for epithelial cell adhesion molecule (EpCAM), a pan-epithelial cell marker. This method yielded 2-3 3 10 6 ATII cells/mouse lung and 1-2 3 10 4 mTECs/trachea that were highly pure (.98%) and viable (.98%).Using these preparations, we found that both ATII cells and mTECs expressed the Lyn tyrosine kinase, which is best studied as an inhibitory kinase in hematopoietic cells. However, we found little or no expression of Syk in either ATII cells or mTECs, which is in contrast to earlier published reports. Both cell types expressed C-type lectin receptors, anaphylatoxin receptors, and various Toll-like receptors (TLRs). In addition, stimulation of ATII cells with TLR ligands led to secretion of various cytokines and chemokines.Interestingly, lyn 2/2 ATII cells were hyperresponsive to TLR3 stimulation, suggesting that, as in hematopoietic cells, Lyn might be playing an inhibitory role in ATII cells. In conclusion, the improved isolation method reported here, along with expression profiles of various immune defense proteins, will help refocus investigations of immune-related signaling events in pulmonary epithelium.

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“…Sustained Kras activity stimulates the self-renewal of AT2 cells, which eventually leads to abnormal tissue growth (1). An AT2 gene-expression signature has been reported (6,7), but it is unclear how the identity of AT2 cells is specified and maintained.…”

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Transcription factor Etv5 is essential for the maintenance of alveolar type II cells

Zhang¹,

Newton²,

Kummerfeld³

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

107

112

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Alveolar type II (AT2) cell dysfunction contributes to a number of significant human pathologies including respiratory distress syndrome, lung adenocarcinoma, and debilitating fibrotic diseases, but the critical transcription factors that maintain AT2 cell identity are unknown. Here we show that the E26 transformation-specific (ETS) family transcription factor Etv5 is essential to maintain AT2 cell identity. Deletion of Etv5 from AT2 cells produced gene and protein signatures characteristic of differentiated alveolar type I (AT1) cells. Consistent with a defect in the AT2 stem cell population, Etv5 deficiency markedly reduced recovery following bleomycin-induced lung injury. Lung tumorigenesis driven by mutant KrasG12D was also compromised by Etv5 deficiency. ERK activation downstream of Ras was found to stabilize Etv5 through inactivation of the cullin-RING ubiquitin ligase CRL4 COP1/DET1 that targets Etv5 for proteasomal degradation. These findings identify Etv5 as a critical output of Ras signaling in AT2 cells, contributing to both lung homeostasis and tumor initiation.A lveolar type II (AT2) cells are a stem cell population that self renews and differentiates into alveolar type I (AT1) cells during lung homeostasis or in response to injury (1, 2). They also give rise to lung adenocarcinoma induced by oncogenic Kras (1, 3-5). AT2 cells are derived from bipotent progenitor cells at the sacculation stage (1). In the adult lung, mature AT2 cells also secrete surfactant phospholipids to maintain normal alveolar function. Sustained Kras activity stimulates the self-renewal of AT2 cells, which eventually leads to abnormal tissue growth (1). An AT2 gene-expression signature has been reported (6, 7), but it is unclear how the identity of AT2 cells is specified and maintained.Given that Ras/MAPK signaling is essential for the self-renewal of AT2 cells and for the initiation of lung adenocarcinoma, identification of the transcription factors that are activated by Ras in AT2 cells is a step toward defining the transcriptional programs underlying AT2 stemness. The PEA3 subgroup of the ETS family of transcription factors, comprised of ETS transcription variants 1, 4, and 5 (Etv1, Etv4, and Etv5) (8), is known to be engaged by Ras/MAPK signaling. Etv4 and Etv5 are expressed in distal lung epithelium during development and are involved in branching morphogenesis and epithelial cell differentiation (9-11). Alveolar epithelial cells in the adult lung also express Etv5 (7, 12), but a critical role for Etv5 in this setting remains elusive.PEA3 transcription factors are also implicated in tumorigenesis. Overexpression of ETV1 or ETV4 has been linked to prostate cancer (13,14), and stabilization of ETV1 by mutant, active tyrosine kinase receptor KIT is thought to drive an oncogenic program in gastrointestinal stromal tumors (GIST) (15). ETV1, ETV4, and ETV5 are labile proteins because of their regulation by the ubiquitin-proteasome system. In GIST, ETV1 protein is stabilized by activation of the Ras/MAPK pathway through an unkno...

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Surfactant Protein–C Chromatin-Bound Green Fluorescence Protein Reporter Mice Reveal Heterogeneity of Surfactant Protein C–Expressing Lung Cells

Cited by 57 publications

References 45 publications

Type 2 alveolar cells are stem cells in adult lung

Type 2 alveolar cells are stem cells in adult lung

Immune Defense Protein Expression in Highly Purified Mouse Lung Epithelial Cells

Transcription factor Etv5 is essential for the maintenance of alveolar type II cells

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